Rhodamine ulex europaeus agglutinin 1

Cross sections (5-μm) of artery grafts were stained with Elastica van Gieson (EVG) as well as H&E and used to quantify the luminal and intimal areas using ImageJ (NIH) for morphometry. The intimal area was defined as bound by internal elastic lamina and the lumen. The EC lining of grafts were identified by Rhodamine ulex europaeus agglutinin 1 (Vector Laboratories) and/or CD31 expression, which yielded equivalent results, and quantified by tracing the circumference of lumen using ImageJ. Graft-infiltrating human lymphocytes were quantified by staining for human CD45RO⁺ (BD Biosciences) or CD3⁺ (Dako), which yielded equivalent results, by indirect immunofluorescence (donkey anti-rabbit or donkey anti-mouse AlexaFluor-594, Invitrogen). The numbers of intimal CD45RO⁺ were counted in two cross sections per graft. All sections were mounted on slides using mounting medium (Pro-Long Gold, Invitrogen) and examined by microscopy with an Axiovert 200M microscope (Zeiss).